title: Unusual transglycosylation activity of Flavobacterium meningosepticum endoglycosidases enables convergent chemoenzymatic synthesis of core fucosylated complex N-glycopeptides.
abstract: Structurally well defined, homogeneous glycopeptides and glycoproteins are indispensable tools for functional glycomics studies. By screening of various endo-β-N-acetylglucosaminidases through the use of appropriate synthetic donor and acceptor substrates, we have found that the Flavobacterium meningosepticum endo-β-N-acetyl-glucosaminidases (GH family 18), including Endo-F2 and Endo-F3, were able to glycosylate α-1,6-fucosylated GlcNAc derivative to provide natural, core-fucosylated complex-type N-glycopeptides. The Endo-F2 and Endo-F3 were efficient for transferring both sialylated and asia-lylated glycans and were highly specific for an α-1,6-fucosylated GlcNAc-peptide as acceptor for transglycosylation, showing only marginal activity with non-fucosylated GlcNAc-peptides. In contrast, we found that the commonly used endoglycosidases such as Endo-A and Endo-M, which belong to GH family 85, were unable to take α-1,6-fucosyl-GlcNAc derivative as acceptors for transglycosylation. The novel activity of Endo-F2 and Endo-F3 was successfully applied for a highly convergent chemoenzymatic synthesis of a full-sized CD52 glycopeptide antigen carrying both terminal sialic acid and core fucose. This is the first report on endoglycosidases that are able to glycosylate α-1,6-fucosylated GlcNAc derivatives to form natural core-fucosylated glycopeptides.
authors.affiliation: Institute of Human Virology and Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland 21201 (USA), Fax: (+1) 410-706-4694.
authors.forename: Wei
authors.initials: W
authors.lastname: Huang
authors.forename: Jie
authors.initials: J
authors.lastname: Li
authors.forename: Lai-Xi
authors.initials: LX
authors.lastname: Wang
mesh_terms: D045247:Chryseobacterium / Q000201:enzymology*; D005643:Fucose / Q000737:chemistry* / Q000378:metabolism; D006023:Glycoproteins / Q000096:biosynthesis*; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006031:Glycosylation; D010080:Oxazoles / Q000737:chemistry
chemical_list: D006023:Glycoproteins; D010080:Oxazoles; C064210:oxazolidine; D005643:Fucose; D006026:Glycoside Hydrolases